Team:Chiba/Calendar-Home/26 August 2008
From 2008.igem.org
(Difference between revisions)
(→Team:Communication) |
(→Team:Output) |
||
Line 199: | Line 199: | ||
<tr> | <tr> | ||
<td>DNA tamplate</td> | <td>DNA tamplate</td> | ||
- | <td>1</td><td>1</td><td>1</td><td>1</td | + | <td>1</td><td>1</td><td>1</td><td>1</td><td>1</td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
Line 211: | Line 211: | ||
<tr> | <tr> | ||
<td>TOTAL</td> | <td>TOTAL</td> | ||
- | <td> | + | <td>6μl</td><td>6μl</td><td>6μl</td><td>6μl</td><td>6μl</td> |
</tr> | </tr> | ||
</table> | </table> | ||
[[Team:Chiba/protocol/digestion|Digestion]] | [[Team:Chiba/protocol/digestion|Digestion]] | ||
- | *vector:[http://partsregistry.org/Part:BBa_F2620 BBa_F2620] | + | *vector:[http://partsregistry.org/Part:BBa_F2620 BBa_F2620]① |
<table width="250" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000"> | <table width="250" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000"> | ||
<tr> | <tr> | ||
<td width="257">Sample No.</td> | <td width="257">Sample No.</td> | ||
- | <td> | + | <td>①</td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
Line 245: | Line 245: | ||
<tr> | <tr> | ||
<td>TOTAL</td> | <td>TOTAL</td> | ||
- | <td> | + | <td>130μl</td> |
</tr> | </tr> | ||
</table> | </table> | ||
- | *insert:[http://partsregistry.org/Part:BBa_J52008 BBa_J52008] | + | *insert:[http://partsregistry.org/Part:BBa_J52008 BBa_J52008]① |
+ | :[http://partsregistry.org/Part:BBa_E2030 BBa_E2030]② | ||
<table width="250" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000"> | <table width="250" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000"> | ||
<tr> | <tr> | ||
<td width="257">Sample No.</td> | <td width="257">Sample No.</td> | ||
- | <td> | + | <td>①</td><td>②</td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
Line 282: | Line 283: | ||
<tr> | <tr> | ||
<td>TOTAL</td> | <td>TOTAL</td> | ||
- | <td> | + | <td>40μl</td><td>40μl</td> |
</tr> | </tr> | ||
</table> | </table> | ||
Line 324: | Line 325: | ||
<tr> | <tr> | ||
<td>TOTAL</td> | <td>TOTAL</td> | ||
- | <td> | + | <td>20μl</td><td>20μl</td><td>20μl</td> |
</tr> | </tr> | ||
</table> | </table> |
Revision as of 15:18, 24 October 2008
25 August 2008 <|> 27 August 2008
Contents |
Laboratory work
Team:Input
no work
Team:Communication
- (8/25)--->Gel Check
|
|
|
--->Gel Extract
- --->Zymo Clean
- eluted with following amount of elution buffer
- 5μl
- 5μl
- 27μl
- --->Gel Check
|
- --->SAP
Sample No. 3 Sample DNA(μL) 26 SAP(μL) 1 SAP Buffer(μL) 3 TOTAL(μL) 30
- --->Zymo Clean
- Sample No.3 -->9μl
- --->Gel Check
Sample No.3 1 Loading Dye(μL) 1 dH2O(μL) 4 TOTAL(μL) 6 - Sample No.3 -> OK ・・・30ng?
- --->Ligation
Sample No. (1) (2) (3) (4) (5) insert①C0178(μL) 1 - - 1 - insert②C0170(μL) - 1 - - 1 vector③J04500(μL) - - 2 2 2 ligase(μL) 1 1 1 1 1 Buffer(μL) 2 2 2 2 2 dH2O(μL) 6 6 5 4 4 TOTAL(μL) 10 10 10 10 10
- --->Transformation
- insert①BBa_C0178
- insert②BBa_C0170
- vector③BBa_J04500
- insert①BBa_C0178+vector③BBa_J04500
- insert②BBa_C0170+vector③BBa_J04500
Team:Output
Sample No. ① ② ③ ④ ⑤ DNA tamplate 1 1 1 1 1 Loading Dye 1 1 1 1 1 dH2O 4 4 4 4 4 TOTAL 6μl 6μl 6μl 6μl 6μl - vector:BBa_F2620①
Sample No. ① DNA tamplate 100 SpeⅠ 2 PstⅠ 2 BSA(x10) 13 Buffer2 13 TOTAL 130μl - insert:BBa_J52008①
Sample No. ① ② DNA tamplate 30 30 XbaⅠ 1 1 PstⅠ 1 1 DpnⅠ 1 1 Buffer3 4 4 BSA(x10) 4 4 TOTAL 40μl 40μl
-->37℃ 3hour
-->SAP
-->37℃ 30min
-->65℃ 15min
-->Zymo Clean- vector:BBa_F2620 insert:BBa_J52008①
- vector:BBa_F2620 insert:BBa_E2030②
- Negative control:BBa_F2620③
Sample No. ① ② ③ vector 5 5 5 inser 10 7.5 0 Ligase Buffer 4 4 4 Ligase 1 1 1 dH2O 0 2.5 10 TOTAL 20μl 20μl 20μl -->RT 2hour
-->Transformation(XL10G)
-->37℃ over night
-->colony PCR
-->Mini prep
- --->Zymo Clean